Analysis of Zuojinwan Based on Ancient Literature / 中国实验方剂学杂志

Zuojinwan originated from Danxi′s Experiential Therapy （《丹溪心法》） in the Yuan dynasty. It is a representative prescription for the treatment of liver fire invading stomach syndrome， and is also one of the typical prescriptions of the anti-adjuvant method of traditional Chinese medicine （TCM）. In this paper， the method of bibliometrics was used to systematically sort out the ancient books of Zuojinwan， and 729 relevant literature data were obtained. After certain retrieval and screening， 57 ancient books of TCM were finally obtained. The statistics and analysis were carried out from the aspects of prescription source， historical evolution， composition， functions， evolution of prescription meaning， prescription dose， and preparation and usage of Zuojinwan. It was found that Zuojinwan was composed of Coptis chinensis rhizoma and Euodia rutaecarpa fructus in a ratio of 6∶1. It was mainly used for the treatment of liver fire invading stomach syndrome. The symptoms included pain in chest and hypochondrium， vomiting and bitter mouth， noisy acid-swallowing， red tongue coating yellow， and pulse string number. Later medical records recorded that Zuojinwan was mostly consistent with the original prescription. It mainly treated various diseases caused by liver fire， including left by liver fire， including left hypochondriac pain， swallowing acid and vomiting acid， tendon hernia and lump， epigastric pain， bitter mouth pulse string， head pain， diarrhea， gonorrhea， cold and hot， abdominal pain， alcohol wet yellowing， silence of oral dysentery and so on. There was little controversy in the analysis of relevant prescriptions. In the past dynasties， pills was mainly used， which was consistent with the original prescription. In modern times， it is mainly water flooding for pills or steamed cakes for pills， warm boiling water to serve 6 g， taking 2-3 g per time， the history is basically the same. In this paper， through the excavation， collation and systematic analysis of the ancient literature of TCM that recorded Zuojinwan， we hope to provide the literature basis for the development， inheritance and utilization of this famous classical formulas.

Establishment of Material Benchmark Characteristic Spectrum of Houpu Qiwutang / 中国实验方剂学杂志

Objective:To establish a characteristic spectrum to reflect the efficacy of Houpo Qiwutang. Method:Based on the correlation between the efficacy and the pharmacological action of each herb in prescription，the target substances of characteristic map were screened. The extraction solvent，detection wavelength and gradient of the active ingredients were optimized. Peak assignment was made by comparing individual drugs. Q-TOF was used to infer the molecular formula of each peak in the characteristic atlas，and the reference substance was identified by the reference substance. The reference substance was screened out according to the correlation of main efficacy and medicine. Result:The characteristic spectrum of material standard of Houpu Qiwutang was established. Five of the seven herbal medicines were attributed. Nine characteristic peaks were selected and identified by Q-TOF as glycyrrhizin，including naringin，neohesperidin，ammonium glycyrrhizinate，rhein，honokiol，magnolol. According to the main efficacy of Houpo Qiwutang，neohesperidin was selected as reference substances. According to the separation of characteristic peaks and the retention time，the mark peak of the characteristic spectrum was determined. Conclusion:The characteristic spectrum of the material basis of Houpo Qiwutang was established by selecting the characteristic peaks and controlling the key components. This method not only reflects the situation of all the effective chemical components，but also focuses on the control of the key efficacy，so as to provide a theoretical basis for the subsequent development and quality control of Houpu Qiwu Tang.

Correlation Analysis of HPLC Fingerprint of Pieces, Standard Decoction, Intermediates and Dispensing Granules of Rehmanniae Radix Praeparata / 中国实验方剂学杂志

Objective::To establish HPLC fingerprint spectra of the pieces, standard decoction, intermediates, dispensing granules of Rehmanniae Radix Praeparata, and assess the quality correlation among them, then to evaluate the scientificity and rationality of preparation process based on the yields of dry extract and the transfer rate of acteoside. Method::Fingerprints of several batches of the pieces, standard decoction, intermediates and dispensing granules of Rehmanniae Radix Praeparata were detected by HPLC, and the content of acteoside was determined according to the method of ChP 2015.The fingerprint chromatographic separation was carried out on Phenomenex Luna 100A C18(2) chromatographic column (4.6 mm×250 mm, 5 μm). The mobile phase was acetonitrile-0.1% phosphoric acid for gradient elution, with a flow rate of 1 mL·min-1, and the detection wavelength was 330 nm. At the same time, the correlation analysis of quality transmission during the preparation of dispensing granules was carried out based on the yields of dry extract and the transfer rates of acteoside. Result::The contents of acteoside pieces, standard decoction and intermediates were basically consistent. The yield of dry extracts of intermediates and dispensing granules, and the transmission rate of acteoside were all within the range of standard decoction, and basically consistent with standard decoction. There were 7 common peaks in all fingerprint spectra of 17 batches of pieces, 17 batches of standard decoction, 10 intermediates and 10 dispensing granules of Rehmanniae Radix Praeparata, with a good correlation. The 13 main chromatographic peaks in the dispensing granules were identified by UPLC-Q-TOF-MS analysis, and 4 of the 7 fingerprint common peaks were identified as 5-hydroxymethyl furfural, acteoside, isoacteoside and martynoside. Conclusion::The main chemical constituents of Rehmanniae Radix Praeparata pieces, standard decoction, intermediates and dispensing granules are basically identical. The established HPLC fingerprint method can be used for the quality control of preparation process of Rehmanniae Radix Praeparata dispensing granules.

Clinical characteristics of cytomegalovirus positive Posner-Schlossman syndrome and the effect of 20g/L Ganciclovir eye drops intervention / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To compare the clinical characteristics of cytomegalovirus positive and negative patients with Posner-Schlossman syndrome(PSS), and the clinical efficacy and short-term recurrence of 20g/L ganciclovir eye drops in local treatment of patients with CMV positive PSS were evaluated. <p>METHODS: Differences between CMV positive(86 cases, 86 eyes)and CMV negative(60 cases, 60 eyes)PSS patients were compared. General information, ocular parameters and distribution, IFN-γ and IL-4 levels in aqueous humor, clinical efficacy and recurrence within 1a were compared between the conventional group(30 cases, 30 eyes)and the experimental group(56 cases, 56 eyes).<p>RESULTS: Among the 146 PSS patients included, the CMV positive rate was 58.9%. The average intraocular pressure and the difference in the number of corneal endothelial cells between the onset eye and the contralateral eye in CMV positive patients were significantly increased, and the number of corneal endothelial cells in the onset eye was significantly decreased(<i>P</i><0.05). After treatment, all ocular parameters and distribution in the experimental group were significantly better than those in the conventional group(<i>P</i><0.05), IFN-γ level in the experimental group was significantly lower than that in the conventional group, and IL-4 level was significantly higher than that in the conventional group(<i>P</i><0.05). Compared with the conventional group, the experimental group had a high total effective rate, a short cure time, a low recurrence rate within 1a and a long recurrence interval, with statistically significant differences(<i>P</i><0.05).<p>CONCLUSION: Patients with CMV positive PSS had higher intraocular pressure and greater damage to corneal endothelial cells than PSS patients with CMV negative. 20g/L ganciclovir eye drops can effectively control intraocular pressure and inflammation by resisting CMV infection, with significant clinical efficacy and not easy to relapse in the short term.

HPLC Fingerprint and Chemical Constituents of Menthae Haplocalycis Herba Standard Decoction / 中国实验方剂学杂志

Objective: To establish an HPLC fingerprint of Menthae Haplocalycis Herba standard decoction,in order to identify the main chemical component of common peaks,and determine the content of rosmarinic acid. Method: The chromatographic fingerprints of 10 batches of Menthae Haplocalycis Herba standard decoction from different areas were determined,and the chromatographic separation was carried out on Kromasil C18 (4.6 mm×250 mm,5 μm) at the temperature of 30℃. The mobile phase was acetonitrile and 0.1%formic acid solution for gradient elution,with a flow rate of 1 mL·min-1,and the detection wavelength was 330 nm. The 10 batches of fingerprints were imported into Similarity Evaluation System for Chromatographic Fingerprint of TCM (2012.130723) for chromatographic peak matching, the reference fingerprint was established with the average method,and the similarities of 10 batches were evaluated. Result: The HPLC fingerprint of Menthae Haplocalycis Herba showed 13 common peaks. The similarities of 10 batches from different areas were all more than 0.90.At the same time,9 common peaks of the fingerprint were identified by using Q-TOF-MS spectrometry. Rosmarinic acid content was also determined by using the HPLC fingerprint method. Conclusion: The method is simple,rapid and accurate,with a good reproducibility, and can be used to rapidly and effectively evaluate the quality of Menthae Haplocalycis Herba standard decoction and lay a foundation for the quality control of Menthae Haplocalycis Herba formula granules.

Treatment of type 2 diabetic peripheral neuropathy patients of qi-yin deficiency complicated phlegm-dampness blocking collaterals syndrome by internal application of qigui mixture and external application of qigui huoxue lotion: a clinical study / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy of internal application of Qigui Mixture (QM) and external application of Qigui Huoxue Lotion (QHL) in treating type 2 diabetic peripheral neuropathy (DNP) patients of qi-yin deficiency complicated phlegm-dampness blocking collaterals syndrome (QYD-PDBCS), and to primarily discuss its mechanism.</p><p><b>METHODS</b>Totally 62 DPN patients of QYD-PDBCS were randomly assigned to the treatment group (31 cases) and the control group (31 cases). All patients received routine comprehensive therapy. Patients in the control group took Mecobalamine Tablet, 500 microg each time, 3 times per day. Patients in the treatment group additionally took QM, 200 mL per day, twice daily. Besides, they had foot bath in QHL 10 - 15 min every evening for 3 months. The efficacy was assessed by Chinese medical symptom integrals and Toronto clinical scoring system (TCSS) before treatment, 2 and 3 months after treatment. The nerve conduction velocity was determined; the serum levels of total antioxidant capacity (T- AOC), malondialdehyde (MDA), and superoxide dismutase (SOD) were detected 2 and 3 months after treatment.</p><p><b>RESULTS</b>The total effective rates of Chinese medical symptom integrals and TCSS score were obviously higher in the treatment group than in the control group (P < 0.05). The nerve conduction velocity was significantly improved in the treatment group, when compared with before treatment (P < 0.01). There was statistical difference in the nerve conduction velocity difference of right median nerve motor branch, bilateral tibial nerve motor branches, bilateral common peroneal nerve motor branches, bilateral ulnar nerve sensory branches, and left tibial nerve sensory branch (P < 0.05). Compared with before treatment, serum levels of T-AOC and SOD significantly increased, and the level of MDA decreased significantly in the treatment group after 2 and 3 months of treatment (P < 0.01). But only the SOD level increased significantly in the control group (P < 0.01). There was no statistical difference in increased T-AOC level between the two groups after 2 months of treatment (P > 0.05), but there was statistical difference in increased SOD level and decreased MDA level (P < 0.05). There was statistical difference in increased T-AOC and SOD levels and decreased MDA level between the two groups after 3 months of treatment (P < 0.05). No adverse reaction occurred during the therapeutic course.</p><p><b>CONCLUSIONS</b>The internal application of QM and external application of QHL combined with Mecobalamine in treating DPN was safe and effective, with more significant efficacy than using Mecobalamine alone. Its mechanism might be associated with resistance to oxidative stress.</p>

Expression of Notch signal molecules in human lymphoma cells and its significance / 中国实验血液学杂志

The study was aimed to investigate the expression of Notch signal molecules in human lymphoma cells and its significance. Raji, Maver, Z138 and Jurkat cell lines were used in the study. RT-PCR was used to determine the expression of Notch signaling molecules in these lymphoma cells. Flow cytometry was used to detect the apoptosis and cell-cycle of the lymphoma cells induced by different concentrations of gamma secretase inhibitor DAPT. CCK-8 was used to detect the proliferation of the lymphoma cells treated by DAPT. The results showed that the expression of Notch molecules in the four cell lines was different. Notch1 and Notch2 were found to be expressed in the four lymphoma cell lines, Notch3 predominantly expressed in Jurkat cells, Notch4 only expressed in Raji cells weakly and Hes1 only expressed in Raji and Jurkat cells. Treatment with DAPT could increase the apoptosis ratio of Raji and Jurkat cells. Besides, DAPT could significantly inhibit the proliferation of Raji and Jurkat cells by inducing the cell cycle arrest in G(1) phase, but the effect of DAPT on Maver and Z138 cells was not obvious. The activity of Notch pathway could be inhibited by DAPT treatment through down-regulating the expression of Notch target gene Hes1. It is concluded that the abnormal expression and activation of Notch signal pathway play an important role in the proliferation of lymphoma cells. Notch may be likely a new target for the therapy of lymphoma.

Effect of chidamide on human B lymphoma cell lines and its mechanisms / 中国实验血液学杂志

This study was aimed to explore the effects of histone deacetylase inhibitor chidamide on the proliferation, apoptosis of B lymphoma cell lines Raji (Burkitt lymphoma), Maver and Z-138 (mantle cell lymphoma) and its mechanisms. Three B lymphoma cell lines were cultured in vitro with different concentrations of chidamide for different time. The cell proliferation was determined by CCK-8 method; the cell apoptosis and mitochondrial membrane potential were analyzed by flow cytometry; the protein levels of histone H3/H4 acetylation in cells and the activity of caspase-3 were detected by Western blot. The results showed that chidamide inhibited the proliferation of 3 B lymphoma cell lines in time- and concentration-dependent manners, especially in Z-138 cell line earlier and faster; chidamide could induce cell apoptosis and decline of mitochondrial membrane potential, which was more sensitive in Maver and Z-138 cells than that in Raji cells. Chidamide could elevate the histone H3/H4 acetylation level in 3 B lymphoma cell lines and the activity of caspase-3 in Maver and Z-138 cells. It is concluded that chidamide can inhibit proliferation of B lymphoma cell lines and promote cell apoptosis, the increase of histone H3/H4 acetylation induced by chidamide, triggering of mitochondrial pathway and activation of caspase-3 may be considered as possible mechanisms.

Overall anatomical features and clinical value of the sacral nerve in high resolution computed tomography reconstruction / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Sacral nerve injury is a common complication of pelvic or sacral fractures. As the sacral nerve courser within the sacrum and has a complex relationship with the surrounding tissues, different parts of the sacral plexus injury have similar clinical symptoms and signs. Since lack of specific imaging technique in the diagnosis of sacral nerve injury, especially on multi-segment, multi-site, how to determine the preoperative location and extent of the sacral nerve injury accurately becomes a concern of the general orthopaedic and images practitioners. This study was conducted to gain an insight into the overall anatomical features of the sacral nerve (SN) on the same slice in high resolution computed tomography (HRCT) reconstruction and to determine the value of this information for the clinical diagnosis of related diseases.</p><p><b>METHODS</b>Fifty healthy volunteers and 30 patients (40 sides) with SN lesions confirmed by surgery were scanned using a 16-slice helical CT scanner (Light Speed, GE, USA). Among the patients, 6 with intervertebral disk hernia (6 sides), 8 with spinal stenosis (12 sides), 11 with pelvic trauma (14 sides), 4 with pelvic malignancies (6 sides), and 1 with sacral vertebral tuberculosis (2 sides). The SN multiplanar reconstruction was performed using a UNIX-based SCD4.1 workstation where the image was set on the same slice. All images were stored in the Digital Imaging and Communications in Medicine format. The display of nerves in different sections was analyzed using a five-graded scale with coordinate curves of each individual score. The overall anatomic features visible on the slice were analyzed and the abnormalities of the lesions were studied.</p><p><b>RESULTS</b>The image of the same slice clearly revealed the shape, running direction, thickness, tension and adjacent anatomy of the S1-S4 nerves. The rank of display rates in different sections was: outward-rotated oblique sagittal > outward-rotated oblique coronal > oblique coronal plane > coronal > sagittal > transverse section. The S5 nerve was partially displayed from the starting point to the segment around the posterior sacral foramen. The overall anatomy of the triangular sacral plexus was only revealed in the oblique outward-rotated sagittal section, while 100% of its individual rami, as well as two or three of the adjacent rami, were displayed from their starting points to the anterior border of the piriformis. The abnormalities included 39 sides of morphological change (97.5%), 38 sides of compression (95.0%), 35 sides of adhesion (87.5%), 32 sides of displacement (80.0%), 34 sides of shrinkage (85.0%), 6 sides of thickening (15.0%), and 2 sides of abruption (5.0%).</p><p><b>CONCLUSIONS</b>The 16-slice CT multiplanar reconstruction was able to reveal the overall anatomic features of the SN on the same slice. The section of reconstruction was a crucial factor in determining the display capability of various sacral nerves. This technology was valuable in the diagnosis and management of related diseases.</p>

Study on the overall anatomical features and clinical value of lumbar nerve root, plexus and branches in CT reconstruction / 中华外科杂志

<p><b>OBJECTIVE</b>To identify the overall anatomical characteristics and the clinical value of the lumbar nerves under CT multiplanar reconstruction.</p><p><b>METHODS</b>Fifty normal subjects and 30 patients with LN diseases (51 sides) were selected, including 10 patients with lumber intervertebral disk hernia, eight patients with spinal stenosis, 5 patients with spondylolisthesis, 1 patient with tethered cord syndrome, 1 patient with lumbar scoliosis, and 5 patients with spinal trauma The 16-slice helical CT (Light Speed, GE Company) was used for scanning the lumbar vertebra with multiplanar reconstruction in Workstation (ADW4.1) with UNIX System in DICOM format. The image was set on the same slice for the overall anatomy and manifestations of the lesions.</p><p><b>RESULTS</b>The same-slice imaging showed the strip-like LN slightly lower than the surrounding muscle in density. Each LN went out of the dural sac at an acute angle. The course of the lumbar plexus and its major branches, including the obturator nerve, femoral nerve and reproductive nerve, and their relations to the adjacent structures were clearly revealed. The percentage of the segments displayed was well associated with the reconstruction angle, with the order being oblique coronal > outward-rotated oblique coronal > oblique sagittal > coronal > sagittal section. The major manifestations of abnormal LN included compression and displacement (50 sides, 98.0%), morphological changes (49 sides, 96.1%), adhesion (41 sides, 80.4%).</p><p><b>CONCLUSIONS</b>The CT multiplanar reconstruction is ideal for the imaging of the overall size, shape, running and tension of the LN root; it is valuable in clinical diagnosis.</p>

Expression, purification and characterization of rat procarboxypeptidase B in Pichia pastoris / 生物工程学报

Carboxypeptidase B is a metalloenzyme, which is widely used for commercial and research purposes. Commercially available CPB purified from porcine or bovine pancreas is very expensive, and is not totally free from other proteases. In order to express the rat proCPB in Pichia pastoris, total RNA extracted from SD rat pancreas cells was reversely transcripted to synthesize cDNA, and the proCPB ORF was synthesized by PCR. After digestion with Xho I and EcoR I , the fragment was inserted into pPIC9, and the recombinant plasmid was named as pPIC9-proCPB. By digestion with Sac I , the lined pPIC9-proCPB was transformed into Pichia pastoris strains GS115 with PEG1000 and integrated into their genomes. In the inducement of methanol, recombinant proCPB was successfully expressed in Pichia pastoris, and could be secreted into the supernatant in the culture. After optimizing the fermentation conditions, a higher production could be obtained when GS115-proCPB was induced in BMGY (pH6.0) at 28CC, with addition of 0.5% casein. The yield of recombinant protein reached 500mg/L, achieving over 94% of total protein in the culture supernatant. The purity of recombinant CPB can reach 96% after two step phenyl sepharose F F purification, and 38% of total protein can obtained after optimizing the pufication method. Comparing to the specific activity 180u/mg of CPB purchased from Sigma, the specific activity of recombinant CPB is 110u/mg. Mass spectrometry analyses showed the mass of the recombinant CPB was 35.1 kD, which is very close to the theory value 35.2 kD. Amino acid sequencing of N-terminal of recombinant CPB further indicated proCPB was expressed successfully and modificated correctly after translation.

Expression and bioactive characterization of bacteriophage lysin gene of Bacillus anthracis in Escherichia coli / 生物工程学报

The lysin gene of Bacillus anthracis-diagnosing bacteriophage, obtained by PCR amplification,was cloned into the Escherichia coli exepression vector pET22b which has been cleaved by EcoR I and Nde I. The recombinant vector pET22b-gamma lysin was verified to be correctly constructed by PCR, sequencing and enzyme digestion, and highly expressed in E. coli BL21 (DE3), which accounted for about 40 percent of total protein in E. coli BL21 (DE3), while in the 5L fermentor the expression level reached 15g/L. After expression, disruption and purification with three-step chromatography, Streamline SP, SP HP and Sephacryl S-100, the recombinant gamma lysin was finally obtained with purity of higher than 95 percent as determined by gel scan. The final yield following SP HP was 19.1 percent, with a greater-than-350-fold increase in specific activity. The pure enzyme has been shown active to Bacillus anthracis, and not to E. coli, Bacillus subtilis and Bacillus cereus. Its specific activity was about 1400 u/mg.

Expression of fusion protein of parathyroid hormone and transferrin N-terminal half-molecule in Pichia pastoris / 生物工程学报

The fused gene (PTH-TFN) of parathyroid hormone (PTH) gene and transferring N-terminal half-molecule (TFN) gene was amplified by multiple PCR and inserted into pPIC9 vector. The recombinant plasmid pPIC9-PTH-TFN was transformed into Pichia pastoris GS115 by PEG. After methanol induction, the target protein was expressed in fermentation supernatant at high level. The fused protein PTH-TFN with purity being higher than 95% was finally obtained after purification through two-step chromatography: SP Sepharose Fast Flow and Phenyl Sepharose Fast Flow. Western blot analysis and adenylate cyclase assay proved that the fused protein exhibited the bioactivity to stimulate cAMP synthesis and the ability to bind Fe3+ in the Fe3+ saturation study as the recombinant TFN did indicating that TFN could be used as the transcellar carrier of PTH.

High expression and characterization of human parathyroid hormone in Escherichia coli / 生物工程学报

Human parathyroid hormone (hPTH) was highly expressed in Escherichia coli by inserted the synthesized whole hPTH cDNA into the vectors pBV220 and pET22b. After expression and disruption, the purified product was acquired through cation exchange chromatography and reverse phase chromatography. From the results of N-terminal sequencing and MALDI-TOF-MS analysis the recombiant prtein was indentified as intact hPTH. In in vitro Bioassays the recombinant hPTH stimulated adenylate cyclase as the standard did. In ovariectomized rats the recombinant hPTH markedly increased the femoral bone mass and bone mineral density.

Features and clinic values of normal lumbar nerve root anatomy with CT on multiple plane reconstruction techniques at the same slice / 中华放射学杂志

Objective To explore features and clinic values of LNR anatomy with multiple planar reconstruction techniques with 16-slice spiral CT at the same slice.Methods The lumbar vertebrae with normal adults of 55 cases and 23 cases with abnormal ENR caused by 8 cases with protrusion of lumbar disc, 5 cases with spinal stenosis,4 cases with malignant tumor,5 cases with trauma and 1 case with lumbar TB confirmed by operation were scanned with 16-slice spiral CT made in American GE company in routine posture of the lumbar vertebrae,reconstructed LNR with UNIX system in workstation (ADW 4.1),and analyzed their normal and abnormal anatomic manifestations at the same slice.Results All of LNR can symmetrically showed on oblique and coronal planes according to different segments:one segment from L1 to L5(55,100% ),two segments: from L1 to L2,L2 to L3 and L3 to L4(55,100% ),three segments: from L1 to L3 (49,88%),from L2to L4(46,84% )and from L3 to L5(20,36% ),four segments: from L1 to L4 (15,27% )and five segments:(8,15% ),respectively.Each LNR,including their whole shapes of passage from starting to end,direction,size,shape,tension and peripheral relationship and so on can showed clearly on oblique and coronal planes and on other planes. However,the later planes can increase LNR but decreasing numbers of LNR and especially increase very long one LNR reconstruction.Primary manifestation of all diseases can be showed on oppressing along its walking line,meanwhile,20 cases with adhesion, 14 cases with displacement,13 cases atrophy and 9 cases with increasing diameter.Conclusions Image anatomy features of full LNR with 16-slice spiral CT with the multiple plane reconstruction techniques is very ideal ways at the same slice.It is a very valuable way to make diagnosis and treatment of LNR diseases.The concept of"road sing"and showing"at the same slice"of LNR are tried to rise from in order to make foundation for studying their image.